ADAMTS13, BioAssay(TM) ELISA Kit (Human)

ADAMTS13 is a zinc-containing metalloprotease enzyme. The ADAMTS13 gene is mapped to chromosome 9q34 by genomic sequence analysis. It is reported that a metal-containing proteolytic enzyme (metalloprotease) in normal plasma cleaves the peptide bond between tyrosine at position 842 and methionine at position 843 in monomeric subunits of von Willebrand factor, thereby degrading the large multimers. And it is confirmed that the ADAMTS13 gene encodes the von Willebrand factor-cleaving protease (VWFCP). ADAMTS13 is secreted in blood and degrades large vWf multimers, decreasing their activity. Sample Type: Cell culture supernates, serum and plasma (heparin, EDTA, citrate). Intended Use: For quantitative detection of human ADAMTS13 in cell culture supernates, serum and plasma(heparin, EDTA, citrate). Sensitivity: <20pg/ml, Range: 0.78-50ng/ml, Specificity: Natural and recombinant human ADAMTS13, Test Principle: This human ADAMTS13 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for ADAMTS13 has been precoated onto 96-well plates. Standards(CHO) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for ADAMTS13 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human ADAMTS13 amount of sample captured in plate. Kit Components: 96-well plate precoated with human ADAMTS13 antibody, 1, Lyophilized recombinant human ADAMTS13 standard, 50ng/tube×2, Biotinylated human ADAMTS13 antibody, 130ul (dilution 1:100), Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100), Sample diluent buffer, 30ml, Antibody diluent buffer, 12ml, ABC diluent buffer, 12ml, TMB color developing agent, 10ml, TMB stop solution, 10ml, Storage and Stability: Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.